scholarly journals Relationship between the Antifungal Susceptibility Profile and the Production of Virulence-Related Hydrolytic Enzymes in Brazilian Clinical Strains ofCandida glabrata

2017 ◽  
Vol 2017 ◽  
pp. 1-10 ◽  
Author(s):  
Maria Helena Galdino Figueiredo-Carvalho ◽  
Lívia de Souza Ramos ◽  
Leonardo Silva Barbedo ◽  
Jean Carlos Almeida de Oliveira ◽  
André Luis Souza dos Santos ◽  
...  

Candida glabratais a facultative intracellular opportunistic fungal pathogen in human infections. Several virulence-associated attributes are involved in its pathogenesis, host-pathogen interactions, modulation of host immune defenses, and regulation of antifungal drug resistance. This study evaluated the in vitro antifungal susceptibility profile to five antifungal agents, the production of seven hydrolytic enzymes related to virulence, and the relationship between these phenotypes in 91 clinical strains ofC. glabrata. AllC. glabratastrains were susceptible to flucytosine. However, some of these strains showed resistance to amphotericin B (9.9%), fluconazole (15.4%), itraconazole (5.5%), or micafungin (15.4%). Overall,C. glabratastrains were good producers of catalase, aspartic protease, esterase, phytase, and hemolysin. However, caseinase and phospholipase in vitro activities were not detected. Statistically significant correlations were identified between micafungin minimum inhibitory concentration (MIC) and esterase production, between fluconazole and micafungin MIC and hemolytic activity, and between amphotericin B MIC and phytase production. These results contribute to clarify some of theC. glabratamechanisms of pathogenicity. Moreover, the association between some virulence attributes and the regulation of antifungal resistance encourage the development of new therapeutic strategies involving virulence mechanisms as potential targets for effective antifungal drug development for the treatment ofC. glabratainfections.

2021 ◽  
Vol 14 (7) ◽  
Author(s):  
Shaghayegh Rostami Yasuj ◽  
Maral Gharaghani ◽  
Seyed Sajjad Khoramrooz ◽  
Marjan Salahi ◽  
Ali Keshtkari ◽  
...  

Background: Candidemia is the most common systemic infection in hospitalized patients causing high mortality. Hence, the diagnosis of this infection in the early stage with appropriate antifungal therapy is paramount. Objectives: The study aimed at molecular identification of Candida species isolated from candidemia patients and evaluation of the in vitro antifungal susceptibility patterns of these strains to fluconazole, amphotericin B, and caspofungin. Methods: In the present study, 800 hospitalized patients who were suspected to have candidemia were sampled. Candida species were isolated and identified based on morphological characteristics and PCR-sequencing of the ITS1-5.8S-ITS2 region. Antifungal susceptibility tests for fluconazole, amphotericin B, and caspofungin were performed according to the Clinical and Laboratory Standards Institute protocol M27-A3. Also, clinical data were recorded from the patients' records. Results: Twenty-seven patients among the sample of hospitalized patients were found to have candidemia. A total of 33.3% of candidemia patients were treated with amphotericin B, in which case the mortality rate was 14.8%. The majority of patients (59%) were from the neonatal intensive care unit, and premature birth was the most common underlying condition. Candida albicans (n = 18; 66.6%) was the most common species isolated from blood cultures, followed by C. parapsilosis (n = 7; 25.9%), C. pelliculosa (n = 1; 3.7%), and C. tropicalis (n = 1; 3.7%). Only one C. albicans isolate resistant to fluconazole (minimum inhibitory concentration = 32 µg/mL). Conclusions: Generally, C. albicans has been the most frequent causative agent of candidemia. Resistance to antifungal drugs among candidemia agents was rare. Also, the identification of Candida isolates at the species level with in vitro antifungal susceptibility tests helps manage candidemia patients better and decrease the mortality rate among them.


2013 ◽  
Vol 57 (10) ◽  
pp. 4769-4781 ◽  
Author(s):  
A. Forastiero ◽  
A. C. Mesa-Arango ◽  
A. Alastruey-Izquierdo ◽  
L. Alcazar-Fuoli ◽  
L. Bernal-Martinez ◽  
...  

ABSTRACTCandida tropicalisranks between third and fourth amongCandidaspecies most commonly isolated from clinical specimens. Invasive candidiasis and candidemia are treated with amphotericin B or echinocandins as first-line therapy, with extended-spectrum triazoles as acceptable alternatives.Candida tropicalisis usually susceptible to all antifungal agents, although several azole drug-resistant clinical isolates are being reported. However,C. tropicalisresistant to amphotericin B is uncommon, and only a few strains have reliably demonstrated a high level of resistance to this agent. The resistance mechanisms operating inC. tropicalisstrains isolated from clinical samples showing resistance to azole drugs alone or with amphotericin B cross-resistance were elucidated. Antifungal drug resistance was related to mutations of the azole target (Erg11p) with or without alterations of the ergosterol biosynthesis pathway. The antifungal drug resistance shownin vitrocorrelated very well with the results obtainedin vivousing the model hostGalleria mellonella. Using this panel of strains, theG. mellonellamodel system was validated as a simple, nonmammalian minihost model that can be used to studyin vitro-in vivocorrelation of antifungals inC. tropicalis. The development inC. tropicalisof antifungal drug resistance with different mechanisms during antifungal treatment has potential clinical impact and deserves specific prospective studies.


2017 ◽  
Vol 61 (5) ◽  
Author(s):  
Mojtaba Taghizadeh-Armaki ◽  
Mohammad Taghi Hedayati ◽  
Saham Ansari ◽  
Saeed Mahdavi Omran ◽  
Sasan Saber ◽  
...  

ABSTRACT Aspergillus flavus has been frequently reported as the leading cause of invasive aspergillosis in certain tropical and subtropical countries. Two hundred A. flavus strains originating from clinical and environmental sources and collected between 2008 and 2015 were phylogenetically identified at the species level by analyzing partial β-tubulin and calmodulin genes. In vitro antifungal susceptibility testing was performed against antifungals using the European Committee on Antimicrobial Susceptibility Testing (EUCAST) broth microdilution method. In addition, genotyping was performed using a short-tandem-repeat (STR) assay of a panel of six microsatellite markers (A. flavus 2A, 2B, 2C, 3A, 3B, and 3C), in order to determine the genetic variation and the potential relationship between clinical and environmental isolates. The geometric means of the minimum inhibitory concentrations/minimum effective concentrations (MICs/MECs) of the antifungals across all isolates were (in increasing order): posaconazole, 0.13 mg/liter; anidulafungin, 0.16 mg/liter; itraconazole, 0.29 mg/liter; caspofungin, 0.42 mg/liter; voriconazole, 0.64 mg/liter; isavuconazole, 1.10 mg/liter; amphotericin B, 3.35 mg/liter; and flucytosine, 62.97 mg/liter. All of the clinical isolates were genetically different. However, an identical microsatellite genotype was found between a clinical isolate and two environmental strains. In conclusion, posaconazole and anidulafungin showed the greatest in vitro activity among systemic azoles and echinocandins, respectively. However, the majority of the A. flavus isolates showed reduced susceptibility to amphotericin B. Antifungal susceptibility of A. flavus was not linked with the clinical or environmental source of isolation. Microsatellite genotyping may suggest an association between clinical and environmental strains, although this requires further investigation.


2015 ◽  
Vol 26 (2) ◽  
pp. 99-104 ◽  
Author(s):  
Edimilson Martins de Freitas ◽  
Larissa Cavalcanti Monteiro ◽  
Michelle Bonfim da Silva Fernandes ◽  
Hercílio Martelli Junior ◽  
Paulo Rogério Ferreti Bonan ◽  
...  

This study aimed to evaluate the in vitro antifungal susceptibility of Candida species of head-and-neck-irradiated patients (Group 1), non-institutionalized (Group 2) and institutionalized elders (Group 3) using Etest(r) methodology. Candida was isolated from saliva and presumptively identified by CHROMagar Candida(r), confirmed by morphological criteria, carbohydrate assimilation (API 20C AUX(r)) and genetic typing (OPE 18). The collection was made from 29, 34 and 29 individuals (Groups 1, 2 and 3, respectively) with 67 isolates. Etest(r) strips (ketoconazole, itraconazole, fluconazole, amphotericin B and flucytosine) on RPMI (Roswell Park Memorial Institute) agar, on duplicate, were used to evaluate susceptibility. ATTC (American Type Culture Collection) 10231 (Candida albicans) was used as quality control. Among the 67 isolates of Candida species, most were susceptible to azoles, flucytosine and amphotericin B. None of the isolates showed resistance and dose-dependent susceptibility to amphotericin B. There were nine strains resistant to itraconazole, six to fluconazole and two to ketoconazole and ten dose-dependent, mainly to flucytocine. The highest MIC (minimum inhibitory concentration) to C. albicans, C. tropicalis, C. parapsilosis was 2.671 μg.mL-1, 8.104 μg.mL-1, 4.429 μg.mL-1, all for flucytosine. C. krusei and C. glabrata were associated with higher MIC for azoles and C. glabrata with higher MIC to flucytosine. In summary, susceptibility to all tested antifungal agents was evident. The isolates were more resistant to itraconazole and dose-dependent to flucytosine. A comparison of C. albicans in the three groups showed no outliers. Higher MIC was associated with C. krusei and C. glabrata.


Author(s):  
Abirami Lakshmy Jayachandran ◽  
Radhika Katragadda ◽  
Ravinder Thyagarajan ◽  
Leela Vajravelu ◽  
Suganthi Manikesi ◽  
...  

Oropharyngeal candidiasis is one of the common manifestations seen in cancer patients on cytotoxic therapy and invasion into deeper tissues can occur if not treated promptly. Emergence of antifungal drug resistance is of serious concern owing to the associated morbidity and mortality. The present study aims at evaluation of clinicomycological association and antifungal drug susceptibility among the 180 recruited patients with cancer on chemotherapy and/or radiotherapy with signs or symptoms suggestive of oral candidiasis. Speciation and antifungal susceptibility was done by Microbroth dilution method for fluconazole, Itraconazole, and Amphotericin B as per standard microbiological techniques. Chi-square test was used for statistical analysis (p<0.05was considered statistically significant).Candida albicanswas the predominant species isolated (94) (58%) followed byCandida tropicalis(34) (20.9%). Fluconazole and Itraconazole showed an overall resistance rate of 14% and 14.8%, respectively. All the isolates were susceptible to Amphotericin B. There was a significant association between the presence of dry mouth and isolation ofCandida(p<0.001). Such clinicomicrobiological associations can help in associating certain symptoms with the isolation ofCandida. Species level identification with in vitro antifungal susceptibility pattern is essential to choose the appropriate drug and to predict the outcome of therapy.


2020 ◽  
Vol 64 (4) ◽  
Author(s):  
Fatima Zohra Delma ◽  
Abdullah M. S. Al-Hatmi ◽  
Jochem B. Buil ◽  
Hein van der Lee ◽  
Marlou Tehupeiory-Kooreman ◽  
...  

ABSTRACT We compared MIC test strip (MTS) and Sensititre YeastOne (SYO) methods with EUCAST and CLSI methods for amphotericin B, 5-fluocytosine, fluconazole, voriconazole, and isavuconazole against 106 Cryptococcus neoformans isolates. The overall essential agreement between the EUCAST and CLSI methods was >72% and >94% at ±1 and ±2 dilutions, respectively. The essential agreements between SYO and EUCAST/CLSI for amphotericin B, 5-flucytosine, fluconazole, and voriconazole were >89/>93% and between MTS and EUCAST/CLSI were >57/>75%. Very major error rates were low for amphotericin B and fluconazole (<3%) and a bit higher for the other drugs (<8%).


1997 ◽  
Vol 41 (4) ◽  
pp. 808-811 ◽  
Author(s):  
I Pujol ◽  
J Guarro ◽  
J Sala ◽  
M D Riba

In vitro antifungal susceptibility testing for filamentous fungi remains unstandardized and is unreliable for determining adequate therapy. A study was performed to evaluate the effect of inoculum size (10(2), 10(3), 10(4), and 10(5) conidia/ml), incubation time (48 and 72 h), and temperature (25, 30, and 35 degrees C) on MICs of amphotericin B for Fusarium spp. (20 strains). The inoculum size showed the clearest effect: when the inoculum was varied from 10(2) to 10(5) conidia/ml, the geometric mean MICs showed increases of between 10- and 19-fold in all the combined conditions of temperature and incubation time assayed. Time of incubation had less effect (increases of between two- and threefold in approximately half of the geometric mean MICs), and temperature especially had little effect (the increases were no higher than twofold). The effects of interaction between inoculum size and temperature on MICs were not statistically significant, while the combined effects of inoculum size and time of reading and of time of reading and temperature produced systematic variation in MICs.


2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Saeid Mahdavi Omran ◽  
Maryam Rezaei Dastjerdi ◽  
Maryam Zuashkiani ◽  
Vahid Moqarabzadeh ◽  
Mojtaba Taghizadeh-Armaki

Background.Candida-associated denture stomatitis (CADS) is a common fungal infection in people who wear dentures. The main objective of this study was to make molecular identification of causative agents of CADS and in vitro antifungal susceptibility testing (AFST) in the Iranian patients with denture stomatitis.Methods. A total of 134Candidaspp. were obtained from patients with denture stomatitis. TheCandidaspp. were identified using a polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) involving the universal internal transcribed spacer (ITS1 and ITS4) primers, which were subjected to digestion with MspI and BlnI restriction enzymes. The in vitro antifungal susceptibility ofCandidaspp. to fluconazole (FLC), terbinafine (TRB), itraconazole (ITC), voriconazole (VRC), posaconazole (POS), ketoconazole (KET), amphotericin B (AMB), and caspofungin (CAS) was evaluated using the Clinical and Laboratory Standards Institute M27-A3 and M27-S4 guidelines.Results. Overall,C. albicanswas the most commonly isolated species (n=84; 62.6%), followed byC. glabrata(n=23; 17.2%),C. tropicalis(n=16; 12%), andC. parapsilosis(n=11; 8.2%). Posaconazole had the lowest geometric mean minimum inhibitory concentration (MIC) (0.03 μg/ml), followed by AMB (0.05 μg/ml), ITC (0.08 μg/ml), VRC (0.11 μg/ml), CAS (0.12 μg/ml), KET (0.15 μg/ml), and FLC (0.26 μg/ml).Discussion. Our study showed thatC. albicanswas most prevalent in Iranian patients with CADS and was susceptible to both azoles and amphotericin B. In addition, POS could be an appropriate alternative to the current antifungal agents used for the treatment of CADS, as well as in the treatment of recurrent candidiasis.


2021 ◽  
Vol 7 (4) ◽  
pp. 271
Author(s):  
Andrew M. Borman ◽  
Mark Fraser ◽  
Zoe Patterson ◽  
Michael D. Palmer ◽  
Elizabeth M. Johnson

Mucoromycoses (infections caused by members of the order Mucorales, phylum Mucoromycota [ex-Zygomycota]) are highly destructive, rapidly progressive infections, with dire prognoses especially when they occur in immunocompromised hosts. Current treatment guidelines recommend liposomal formulations of amphotericin B with adjunctive surgery as first line therapy, with the newer triazoles posaconazole or isavuconazole as alternative treatments, or as salvage therapy. Among the many organisms belonging to this order, a limited number of species in the genera Rhizopus, Mucor, Lichtheimia and Rhizomucor are responsible for most cases of human infection. Here, we present the minimum inhibitory concentration data (MICs) for amphotericin B, posaconazole, isavuconazole, itraconazole and voriconazole with a panel of over 300 isolates of the five most common agents of human infection (Lichtheimia corymbifera, Rhizopus arrhizus, R. microsporus, Rhizomucor pusillus and Mucor spp.) determined using the CLSI broth microdilution method. In agreement with previous studies, the most active antifungal drug for all Mucorales was amphotericin B, with MICs within the range that would predict susceptibility with Aspergillus fumigatus. Conversely, MICs for voriconazole against all species tested were high, and above the range associated with clinical efficacy with A. fumigatus. Interestingly, whilst isavuconazole and posaconazole MIC distributions indicated in vitro activity against some members of the Mucorales, activity was species-dependent for both agents. These data underscore the importance of accurate identification of the causative agents of mucoromycosis, coupled with antifungal susceptibility testing of individual isolates, in determining the optimal treatment of infections caused by these aggressive opportunistic human fungal pathogens.


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